E-Notebook

From 2007.igem.org

(Difference between revisions)
(→Tasks Accomplished)
(→Experiments so far)
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=== Experiments so far ===
=== Experiments so far ===
-
#Calibration of the number of colonies obtained at different optical densities.
+
*Calibration of the number of colonies obtained at different optical densities.
-
#Transformation of competent E.coli (strain K12Z1) cells with constructs.
+
*Transformation of competent E.coli (strain K12Z1) cells with constructs.
-
#'''Induction of the construct pLac cfp'''
+
*'''Induction of the construct pLac cfp'''
-
E.coli cells transformed with the construct pLac cfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.  
+
E.coli cells transformed with the construct pLac cfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.
-
 
+
-
The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used
+
The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used.
{| cellpadding="2" cellspacing="3" border="1"
{| cellpadding="2" cellspacing="3" border="1"
!Sample
!Sample
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'''Results:'''
'''Results:'''
-
 
The mean fluorescence of CFP was seen to increase with increasing concentrations of the IPTG.
The mean fluorescence of CFP was seen to increase with increasing concentrations of the IPTG.
 +
 +
*'''Induction of the construct pLac yfp'''
 +
E.coli cells transformed with the construct pLac yfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.
 +
 +
The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used.
 +
{| cellpadding="2" cellspacing="3" border="1"
 +
!Sample
 +
!100mM IPTG (ul)
 +
!1mM IPTG (ul)
 +
!Inoculum volume from LB (ul)
 +
!Volume of  M9 added (ml)
 +
|-align="center"
 +
|pLac yfp (0 uM)
 +
| -
 +
| -
 +
|3 ul
 +
|3 ml
 +
|-align="center"
 +
|pLac yfp (1 uM)
 +
| -
 +
|3 ul
 +
|3 ul
 +
|3 ml
 +
|-align="center"
 +
|pLac yfp (10 uM)
 +
| -
 +
|30 ul
 +
|3 ul
 +
|3 ml
 +
|-align="center"
 +
|pLac yfp (50 uM)
 +
|1.5 ul
 +
| -
 +
|3 ul
 +
|3 ml
 +
|-align="center"
 +
|pLac yfp (100 uM)
 +
|3 ul
 +
| -
 +
|3 ul
 +
|3 ml
 +
|-align="center"
 +
|pLac yfp (1000 uM)
 +
|30 ul
 +
| -
 +
|3 ul
 +
|3 ml
 +
|-align="center"
 +
|K12Z1 (Autoflourescence)
 +
| -
 +
| -
 +
|3 ul
 +
|3 ml
 +
|}
 +
 +
'''Results:'''
 +
The mean fluorescence of YFP was seen to increase with increasing concentrations of the IPTG.
== The Week Ahead ==
== The Week Ahead ==

Revision as of 07:12, 27 June 2007

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The official wiki of the NCBS iGEM 2007 Team
[http://www.ncbs.res.in/ National Centre for Biological Sciences, Bangalore]
Bangalore The Company The Mission e-Notebook


The Bangalore iGEM Journal, 07




Contents

e-notebook

May 07 June 07 July 07
S M T W T F S S M T W T F S S M T W T F S
1 2 3 4 5 1 2 1 2 3 4 5 6 7
6 7 8 9 10 11 12 3 4 5 6 7 8 9 8 9 10 11 12 13 14
13 14 15 16 17 18 19 10 11 12 13 14 15 16 15 16 17 18 19 20 21
20 21 22 23 24 25 26 17 18 19 20 21 22 23 22 23 24 25 26 27 28
27 28 29 30 31 24 25 26 27 28 29 30 29 30

Current Status

Tasks Accomplished

Experiments so far

  • Calibration of the number of colonies obtained at different optical densities.
  • Transformation of competent E.coli (strain K12Z1) cells with constructs.
  • Induction of the construct pLac cfp

E.coli cells transformed with the construct pLac cfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.

The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used.

Sample 100mM IPTG (ul) 1mM IPTG (ul) Inoculum volume from LB (ul) Volume of M9 added (ml)
pLac cfp (0 uM) - - 3 ul 3 ml
pLac cfp (1 uM) - 3 ul 3 ul 3 ml
pLac cfp (10 uM) - 30 ul 3 ul 3 ml
pLac cfp (50 uM) 1.5 ul - 3 ul 3 ml
pLac cfp (100 uM) 3 ul - 3 ul 3 ml
pLac cfp (1000 uM) 30 ul - 3 ul 3 ml
K12Z1 (Autoflourescence) - - 3 ul 3 ml

Cells were allowed to grow till the optical density was in the range of 0.05-0.1 (early exponential phase) and were imaged using a phase contrast microscope.

Results: The mean fluorescence of CFP was seen to increase with increasing concentrations of the IPTG.

  • Induction of the construct pLac yfp

E.coli cells transformed with the construct pLac yfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.

The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used.

Sample 100mM IPTG (ul) 1mM IPTG (ul) Inoculum volume from LB (ul) Volume of M9 added (ml)
pLac yfp (0 uM) - - 3 ul 3 ml
pLac yfp (1 uM) - 3 ul 3 ul 3 ml
pLac yfp (10 uM) - 30 ul 3 ul 3 ml
pLac yfp (50 uM) 1.5 ul - 3 ul 3 ml
pLac yfp (100 uM) 3 ul - 3 ul 3 ml
pLac yfp (1000 uM) 30 ul - 3 ul 3 ml
K12Z1 (Autoflourescence) - - 3 ul 3 ml

Results: The mean fluorescence of YFP was seen to increase with increasing concentrations of the IPTG.

The Week Ahead

The to-do List

Discussions

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